Effects of sequence context on O6-alkylguanine DNA alkyltransferase repair of O6-alkyl-deoxyguanosine adducts

Rebecca Guza, Anthony Pegg, Natalia Tretyakova

Research output: Chapter in Book/Report/Conference proceedingConference contribution

6 Citations (Scopus)

Abstract

The DNA repair protein, O6-alkylguanine DNAalkyltransferase (AGT), specifically recognizes and removes O6-alkyl substituents on guanine, restoring normal guanine and preventing mutagenesis. When AGT binds O6-alkyl-deoxyguanosine (O6-alkyl-dG) containing DNA, the protein interacts with the 1-, N2-, and 7- positions of the guanine lesion. The repair of O6-alkyl-dG lesions is a multi-step process. Following AGT binding to DNA, the adducted nucleotide is flipped into the active site of the protein, and the O6-alkyl substituent is displaced via an SN2-type mechanism, restoring normal guanine and producing alkylated protein. The rates of AGT repair are influenced by DNA sequence context, secondary structure, and alkyl group identity. The relative rates of AGT-mediated repair of O6-alkyl-dG lesions are benzyl > methyl > ethyl ≫ 2-hydroxyethyl >4-(3-pyridyl)-4-oxobutyl. The differences in rates of repair between different alkyl groups and different sequence contexts are not a result of difference in AGT binding and kinetics of nucleotide flipping, since these reaction steps are very fast and are unaffected by DNA sequence. The rate of alkyl transfer is the slowest forward step in the repair of O6-Me-dG and appears to be dependent on the alkyl group identity and is influenced by the local sequence context. AGT repair of O6-alkyl-dG lesions is essential for maintenance of genome integrity, and slow repair of these lesions in specific DNA sequences may contribute to the mutational spectra observed in human cancer.

Original languageEnglish (US)
Title of host publicationStructural Biology of DNA Damage and Repair
PublisherAmerican Chemical Society
Pages73-101
Number of pages29
ISBN (Print)9780841225749
DOIs
StatePublished - Jun 1 2010

Publication series

NameACS Symposium Series
Volume1041
ISSN (Print)0097-6156
ISSN (Electronic)1947-5918

Fingerprint

Deoxyguanosine
DNA
Repair
Guanine
DNA sequences
Proteins
Nucleotides
Mutagenesis
DNA alkyltransferase
Genes
Kinetics

All Science Journal Classification (ASJC) codes

  • Chemistry(all)
  • Chemical Engineering(all)

Cite this

Guza, R., Pegg, A., & Tretyakova, N. (2010). Effects of sequence context on O6-alkylguanine DNA alkyltransferase repair of O6-alkyl-deoxyguanosine adducts. In Structural Biology of DNA Damage and Repair (pp. 73-101). (ACS Symposium Series; Vol. 1041). American Chemical Society. https://doi.org/10.1021/bk-2010-1041.ch006
Guza, Rebecca ; Pegg, Anthony ; Tretyakova, Natalia. / Effects of sequence context on O6-alkylguanine DNA alkyltransferase repair of O6-alkyl-deoxyguanosine adducts. Structural Biology of DNA Damage and Repair. American Chemical Society, 2010. pp. 73-101 (ACS Symposium Series).
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abstract = "The DNA repair protein, O6-alkylguanine DNAalkyltransferase (AGT), specifically recognizes and removes O6-alkyl substituents on guanine, restoring normal guanine and preventing mutagenesis. When AGT binds O6-alkyl-deoxyguanosine (O6-alkyl-dG) containing DNA, the protein interacts with the 1-, N2-, and 7- positions of the guanine lesion. The repair of O6-alkyl-dG lesions is a multi-step process. Following AGT binding to DNA, the adducted nucleotide is flipped into the active site of the protein, and the O6-alkyl substituent is displaced via an SN2-type mechanism, restoring normal guanine and producing alkylated protein. The rates of AGT repair are influenced by DNA sequence context, secondary structure, and alkyl group identity. The relative rates of AGT-mediated repair of O6-alkyl-dG lesions are benzyl > methyl > ethyl ≫ 2-hydroxyethyl >4-(3-pyridyl)-4-oxobutyl. The differences in rates of repair between different alkyl groups and different sequence contexts are not a result of difference in AGT binding and kinetics of nucleotide flipping, since these reaction steps are very fast and are unaffected by DNA sequence. The rate of alkyl transfer is the slowest forward step in the repair of O6-Me-dG and appears to be dependent on the alkyl group identity and is influenced by the local sequence context. AGT repair of O6-alkyl-dG lesions is essential for maintenance of genome integrity, and slow repair of these lesions in specific DNA sequences may contribute to the mutational spectra observed in human cancer.",
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Guza, R, Pegg, A & Tretyakova, N 2010, Effects of sequence context on O6-alkylguanine DNA alkyltransferase repair of O6-alkyl-deoxyguanosine adducts. in Structural Biology of DNA Damage and Repair. ACS Symposium Series, vol. 1041, American Chemical Society, pp. 73-101. https://doi.org/10.1021/bk-2010-1041.ch006

Effects of sequence context on O6-alkylguanine DNA alkyltransferase repair of O6-alkyl-deoxyguanosine adducts. / Guza, Rebecca; Pegg, Anthony; Tretyakova, Natalia.

Structural Biology of DNA Damage and Repair. American Chemical Society, 2010. p. 73-101 (ACS Symposium Series; Vol. 1041).

Research output: Chapter in Book/Report/Conference proceedingConference contribution

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N2 - The DNA repair protein, O6-alkylguanine DNAalkyltransferase (AGT), specifically recognizes and removes O6-alkyl substituents on guanine, restoring normal guanine and preventing mutagenesis. When AGT binds O6-alkyl-deoxyguanosine (O6-alkyl-dG) containing DNA, the protein interacts with the 1-, N2-, and 7- positions of the guanine lesion. The repair of O6-alkyl-dG lesions is a multi-step process. Following AGT binding to DNA, the adducted nucleotide is flipped into the active site of the protein, and the O6-alkyl substituent is displaced via an SN2-type mechanism, restoring normal guanine and producing alkylated protein. The rates of AGT repair are influenced by DNA sequence context, secondary structure, and alkyl group identity. The relative rates of AGT-mediated repair of O6-alkyl-dG lesions are benzyl > methyl > ethyl ≫ 2-hydroxyethyl >4-(3-pyridyl)-4-oxobutyl. The differences in rates of repair between different alkyl groups and different sequence contexts are not a result of difference in AGT binding and kinetics of nucleotide flipping, since these reaction steps are very fast and are unaffected by DNA sequence. The rate of alkyl transfer is the slowest forward step in the repair of O6-Me-dG and appears to be dependent on the alkyl group identity and is influenced by the local sequence context. AGT repair of O6-alkyl-dG lesions is essential for maintenance of genome integrity, and slow repair of these lesions in specific DNA sequences may contribute to the mutational spectra observed in human cancer.

AB - The DNA repair protein, O6-alkylguanine DNAalkyltransferase (AGT), specifically recognizes and removes O6-alkyl substituents on guanine, restoring normal guanine and preventing mutagenesis. When AGT binds O6-alkyl-deoxyguanosine (O6-alkyl-dG) containing DNA, the protein interacts with the 1-, N2-, and 7- positions of the guanine lesion. The repair of O6-alkyl-dG lesions is a multi-step process. Following AGT binding to DNA, the adducted nucleotide is flipped into the active site of the protein, and the O6-alkyl substituent is displaced via an SN2-type mechanism, restoring normal guanine and producing alkylated protein. The rates of AGT repair are influenced by DNA sequence context, secondary structure, and alkyl group identity. The relative rates of AGT-mediated repair of O6-alkyl-dG lesions are benzyl > methyl > ethyl ≫ 2-hydroxyethyl >4-(3-pyridyl)-4-oxobutyl. The differences in rates of repair between different alkyl groups and different sequence contexts are not a result of difference in AGT binding and kinetics of nucleotide flipping, since these reaction steps are very fast and are unaffected by DNA sequence. The rate of alkyl transfer is the slowest forward step in the repair of O6-Me-dG and appears to be dependent on the alkyl group identity and is influenced by the local sequence context. AGT repair of O6-alkyl-dG lesions is essential for maintenance of genome integrity, and slow repair of these lesions in specific DNA sequences may contribute to the mutational spectra observed in human cancer.

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Guza R, Pegg A, Tretyakova N. Effects of sequence context on O6-alkylguanine DNA alkyltransferase repair of O6-alkyl-deoxyguanosine adducts. In Structural Biology of DNA Damage and Repair. American Chemical Society. 2010. p. 73-101. (ACS Symposium Series). https://doi.org/10.1021/bk-2010-1041.ch006